Mouse CD47 Antibody

Catalog # Availability Size / Price Qty
MAB18661-100
MAB18661-SP
Detection of CD47 in HEK293 Human Cell Line Transfected with Mouse CD47 and eGFP by Flow Cytometry.
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Mouse CD47 Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse CD47 in direct ELISAs.
Source
Monoclonal Rat IgG1 Clone # 974222
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived mouse CD47
Gln19-Pro158
Accession # NP_034711
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Flow Cytometry
0.25 µg/106 cells
See below
Blockade of Receptor-ligand Interaction
In a functional ELISA, 0.08-0.8 µg/mL of this antibody will block 50% of the binding of 0.25 µg/mL of Recombinant Mouse CD47 Fc Chimera (Catalog # 1866-CD) to immobilized Recombinant Mouse SIRP alpha /CD172a Fc Chimera (Catalog # 7154-SA) coated at 1 µg/mL (100 µL/well). At 5 µg/mL, this antibody will block >90% of the binding.
 
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry Detection of CD47 antibody in HEK293 Human Cell Line Transfected with Mouse CD47 and eGFP antibody by Flow Cytometry. View Larger

Detection of CD47 in HEK293 Human Cell Line Transfected with Mouse CD47 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with either (A) mouse CD47 or (B) irrelevant protein, and eGFP was stained with Rat Anti-Mouse CD47 Monoclonal Antibody (Catalog # MAB18661) followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113). Quadrant markers were set based on isotype control antibody (Catalog # MAB005, data not shown). View our protocol for Staining Membrane-associated Proteins.

Blockade of Receptor-ligand Interaction CD47 Binding to SIRP  alpha /CD172a Blocked By Mouse CD47 Antibody. View Larger

CD47 Binding to SIRP alpha /CD172a Blocked By Mouse CD47 Antibody. In a functional ELISA, 0.08-0.8 µg/mL of this antibody (green line) will block 50% of the binding of 0.25 µg/mL of Recombinant Mouse CD47 Fc Chimera (orange line, Catalog # 1866-CD) to immobilized Recombinant Mouse SIRPa/CD172a Fc Chimera (Catalog # 7154-SA) coated at 1 µg/mL (100 µL/well). At 5 µg/mL, this antibody will block >90% of the binding.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: CD47

CD47, also known as Integrin‑Associated Protein (IAP) and OA3, is a 40‑60 kDa variably glycosylated atypical member of the immunoglobulin superfamily (1, 2). Mouse CD47 is an integral membrane protein that consists of a 122 amino acid (aa) extracellular domain (ECD) with a single Ig‑like domain, five membrane-spanning regions with short intervening loops, and a 16 aa C‑terminal cytoplasmic tail (3). Alternate splicing of mouse CD47 generates an additional isoform with an insertion of 21 aa following the Ig‑like domain (3). Within the N‑terminal ECD, mouse CD47 shares 63% and 84% aa sequence identity with human and rat CD47, respectively. A portion of the N‑terminal ECD can by shed from smooth muscle cells by MMP-2‑mediated proteolysis (4). The ubiquitously expressed CD47 binds to SIRP family members on macrophages, neutrophils, and T cells (5, 6). These interactions prevent macrophage‑mediated clearance of healthy CD47-expressing cells and promote immune cell transmigration across the vascular endothelium (5‑8). The CD47-SIRP alpha interaction is species specific, and this lack of cross-species interaction has been implicated in xenotransplantation rejection (16). CD47 associates in cis with Fas on T cells and enhances Fas‑mediated apoptosis; its ligation promotes T cell anergy and dampens Th1 immune responses (9‑11). CD47 also associates in cis with Integrins alpha 4 beta 1, alpha V beta 3, alpha 2b beta 3, and alpha 2 beta 1 which can positively or negatively modulate Integrin-mediated function (2, 12). In the vasculature, CD47 binding by Thrombospondin‑1 inhibits the angiogenic and vasorelaxant effects of nitric oxide (2, 13, 14). On dendritic cells and myeloma cells, CD47 ligation by TSP‑1 induces giant cell formation and osteoclast differentiation (15).

References
  1. Sarfati, M. et al. (2009) Curr. Drug Targ. 9:842.
  2. Isenberg, J.S. et al. (2008) Arterioscler. Thromb. Vasc. Biol. 28:615.
  3. Lindberg, F.P. et al. (1993) J. Cell Biol. 123:485.
  4. Maile, L.A. et al. (2008) Mol. Endocrinol. 22:1226.
  5. Oldenborg, P.-A. et al. (2000) Science 288:2051.
  6. Liu, Y. et al. (2002) J. Biol. Chem. 277:10028.
  7. Stefanidakis, M. et al. (2008) Blood 112:1280.
  8. de Vries, H.E. et al. (2002) J. Immunol. 168:5832.
  9. Manna, P.P. et al. (2005) J. Biol. Chem. 280:29637.
  10. Avice, M.-N. et al. (2001) J. Immunol. 167:2459.
  11. Bouguermouh, S. et al. (2008) J. Immunol. 180:8073.
  12. Barazi, H.O. et al. (2002) J. Biol. Chem. 277:42859.
  13. Isenberg, J.S. et al. (2009) Nitric Oxide 21:52.
  14. Isenberg, J.S. et al. (2009) Matrix Biol. 28:110.
  15. Kukreja, A. et al. (2009) Blood 114:3413.
  16. Wang H. et al. (2007) Blood 109:836.
Entrez Gene IDs
961 (Human); 16423 (Mouse); 397042 (Porcine); 478552 (Canine); 102139846 (Cynomolgus Monkey)
Alternate Names
antigen identified by monoclonal 1D8; Antigenic surface determinant protein OA3; CD47 antigen (Rh-related antigen, integrin-associated signal transducer); CD47 antigen; CD47 glycoprotein; CD47 molecule; CD47; IAP; IAPintegrin associated protein; Integrin-associated protein; leukocyte surface antigen CD47; MER6; MER6integrin-associated signal transducer; OA3; Protein MER6; Rh-related antigen

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